Crystalline forms of therapeutic compounds and uses thereof

ABSTRACT

Described herein are crystalline forms of Compound 1 useful for the prevention and treatment of angiogenic ocular conditions; methods of treating a disease comprising; processes for preparing; and kits comprising, the same.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a division of U.S. patent application Ser.No. 16/286,216 filed Feb. 26, 2019, which is a division of U.S. patentapplication Ser. No. 15/694,331 filed Sep. 1, 2017, now U.S. Pat. No.10,253,036, which claims the benefit of U.S. Provisional PatentApplication 62/385,154 filed Sep. 8, 2016. The entire contents of bothof these applications are incorporated by reference herein.

FIELD

This disclosure relates to crystalline forms of a therapeutic compounduseful for treating diseases, including proliferative diseases anddiseases associated with angiogenesis, such as cancer and maculardegeneration.

BACKGROUND

Growth factors play an important role in angiogenesis,lymphangiogenesis, and vasculogenesis. Growth factors regulateangiogenesis in a variety of processes including embryonic development,wound healing, and several aspects of female reproductive function.Undesirable or pathological angiogenesis is associated with diseasesincluding diabetic retinopathy, psoriasis, cancer, rheumatoid arthritis,atheroma, Kaposi's sarcoma, and hemangioma. Angiogenic ocular conditionsrepresent the leading cause of irreversible vision loss in developedcountries. In the United States, for example, retinopathy ofprematurity, diabetic retinopathy, and age-related macular degenerationare the principal causes of blindness in infants, working age adults,and the elderly, respectively. Efforts have been developed to inhibitangiogenesis in the treatment of these conditions.

Therefore, there is a need for new therapeutic compositions for thetreatment of diseases associated with the aberrant signaling of growthfactors and diseases associated with angiogenesis, such as cancer,macular degeneration, and diabetic retinopathy.

SUMMARY

Compound 1 may be useful for the treatment of the proliferative diseasesassociated with angiogenesis, such as angiogenic ocular diseases. Thisdisclosure relates to crystalline forms of Compound 1.

In some embodiments, the crystalline form has an X-ray powderdiffraction (XRPD) pattern with a largest peak at about 10-11 degrees2θ.

Some embodiments include, a crystalline form of Compound 1 having anX-ray powder diffraction (XRPD) pattern with a largest peak at about17-18 degrees 2θ.

Some embodiments include a process for preparing a crystalline form Edescribed herein, comprising mixing Compound 1 and methanol.

Some embodiments include a process for preparing a crystalline form Fdescribed herein, comprising crystallizing Compound 1 in water andisopropanol.

Some embodiments include a pharmaceutical composition comprising thecrystalline form disclosed herein.

Some embodiments include a kit comprising the crystalline form disclosedherein, or the pharmaceutical composition disclosed herein.

Some embodiments include a method of treating a disease comprisingadministering to a subject in need thereof a therapeutically effectiveamount of the crystalline form or a pharmaceutical composition.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts the X-ray powder diffraction (XRPD) pattern ofcrystalline form E.

FIG. 2 depicts an XRPD pattern of crystalline form F.

DETAILED DESCRIPTION

This description relates to crystal or crystalline forms of the compound7-(3-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinazolin-7-yloxy)propyl)-2-oxa-7-azaspiro[3.5]nonane,or a salt thereof, referred to herein as Compound 1 and shown below:

In particular, two crystalline forms of Compound 1, crystalline form Eand crystalline form F, are described herein.

Several X-ray powder diffraction (XRPD) patterns are depicted anddescribed herein. As used herein, the “largest peak” refers to the peakin a diffraction pattern with the highest intensity. As used herein, theterm “major intensity peak” includes any peak having an intensity thatis in the top 20% of the peaks in a particular X-ray powder diffractionpattern.

Crystalline Form E

Crystalline form E has an XRPD pattern with a largest peak at about10-11 degrees two theta (2θ). The diffraction pattern may also have amajor intensity peak at about 15-16 degrees 2θ, about 18-19 degrees 2θ,about 23-23.5 degrees 2θ, and/or about 23.5-24 degrees 2θ, and may haveother major intensity peaks.

Crystalline form E may be a solvate, such as a methanol solvate.

While there are many ways that crystalline form E may potentially beprepared, in some embodiments, crystalline form E may be prepared bystirring a slurry of amorphous of Compound 1 in methanol.

Crystalline Form F

Crystalline form F has an XRPD pattern with a largest peak at about17-18 degrees 2θ (e.g., about 17.65 degrees 2θ). The diffraction patternmay also have a major intensity peak at about 7-8 degrees 2θ (e.g.,about 7.74 degrees 2θ), about 12.5-13.5 degrees 2θ (e.g., about 12.94degrees 2θ), about 14.5-15.5 degrees 2θ (e.g., about 14.87 degrees 2θ),and/or about 26-27 degrees 2θ (e.g., about 26.62 degrees 2θ), and mayhave other major intensity peaks. In some embodiments, crystalline formF has an XRPD pattern as shown in FIG. 2.

Crystalline form F may be a hydrate.

While there are many ways that crystalline form F may be prepared,crystalline form F may be prepared by crystallization of amorphous ofCompound 1 or crystalline form E of Compound 1 from water andisopropanol.

Methods of Treatment

Compound 1 may be used for treating and/or preventing a diseaseassociated with aberrant signaling of a growth factor, such as vascularendothelial growth factor (VEGF). In some embodiments, Compound 1 in acrystalline form may be used to treat a disease associated with abnormalangiogenesis, such as cancer, benign neoplasm, atherosclerosis,hypertension, inflammatory disease, rheumatoid arthritis, maculardegeneration (AMD), choroidal neovascularization, retinalneovascularization, and diabetic retinopathy. In certain embodiments,Compound 1 is used to treat cancer (e.g., an ocular cancer). In certainembodiments, Compound 1 is used to treat macular degeneration.

Compound 1 may also be used to treat or prevent a proliferative disease,such as cancer, ocular disease (e.g., retinopathy, age-related maculardegeneration (AMD), corneal neovascularization, diabetic macular edema,retinal vein occlusion etc.).

The term “ocular disease” or “ocular disorder” includes any eye diseaseand/or disorder. For example, ocular diseases can be disorders of theeyelid, lacrimal system and orbit, disorders of conjunctiva, disordersof sclera, cornea, iris and ciliary body, disorders of choroid,disorders of retina, glaucoma, disorders of optic nerve and visualpathways, ocular neovascularization diseases or disorders, ocularinflammatory diseases, or disorders of ocular muscles. Additionally,ocular disease can also refer to discomfort following injury, surgery,or laser treatment. Diseases and disorders of the eye or ocular diseasesinclude, but are not limited to, retinopathy, diabetic retinopathy,retinal vein occlusion, macular degeneration, age-related maculardegeneration, dry eye syndrome, blepharitis, inflammatory meibomiangland disease, uveitis, allergic conjunctivitis, glaucoma, macularedema, diabetic macular edema, cystoid macular edema, and rosacea (ofthe eye). Dry eye syndrome (DES), otherwise known askeratoconjunctivitis sicca (KCS), keratitis sicca, sicca syndrome, orxerophthalmia, is an eye disease caused by decreased tear production orincreased tear film evaporation commonly found in humans and someanimals.

The term “age-related macular degeneration” or “AMD” includes an oculardisease which usually affects older adults and results in a loss ofvision in the center of the visual field (the macula) because of damageto the retina. It occurs in “dry” and “wet” forms. It is a major causeof blindness and visual impairment in older adults (>50 years).

Macular degeneration can make it difficult or impossible to read orrecognize faces, although enough peripheral vision remains to allowother activities of daily life. The macula is the central area of theretina, which provides the most detailed central vision. In the dry(nonexudative) form, cellular debris called drusen accumulate betweenthe retina and the choroid, and the retina can become detached. In thewet (exudative) form, which is more severe, blood vessels grow up fromthe choroid behind the retina, and the retina can also become detached.It can be treated with laser coagulation, and with medication that stopsand sometimes reverses the growth of blood vessels. Macular degenerationincludes some macular dystrophies affecting younger subjects as well asage-related macular degeneration (AMD or ARMD), which is more commonlyknown. AMD begins with characteristic yellow deposits (drusen) in themacula, between the retinal pigment epithelium and the underlyingchoroid. Most patients with these early changes (referred to asage-related maculopathy) have good vision. Patients with drusen can goon to develop advanced AMD. The risk is considerably higher when thedrusen are large and numerous and associated with disturbance in thepigmented cell layer under the macula. Recent research suggests thatlarge and soft drusen are related to elevated cholesterol deposits andmay respond to cholesterol-lowering agents.

The term “macular edema” refers to the ocular diseases cystoid macularedema (CME) or diabetic macular edema (DME). CME is an ocular diseasewhich affects the central retina or macula of the eye. When thiscondition is present, multiple cyst-like (cystoid) areas of fluid appearin the macula and cause retinal swelling or edema. CME may accompany avariety of diseases such as retinal vein occlusion, uveitis, and/ordiabetes. CME commonly occurs after cataract surgery. DME occurs whenblood vessels in the retina of patients with diabetes begin to leak intothe macula. These leaks cause the macula to thicken and swell,progressively distorting acute vision. While the swelling may not leadto blindness, the effect can cause a severe loss in central vision.

The term “glaucoma” refers to an ocular disease in which the optic nerveis damaged in a characteristic pattern. This can permanently damagevision in the affected eye and lead to blindness if left untreated. Itis normally associated with increased fluid pressure in the eye (aqueoushumor). The term ocular hypertension is used for patients withconsistently raised intraocular pressure (IOP) without any associatedoptic nerve damage. Conversely, the term normal tension or low tensionglaucoma is used for those with optic nerve damage and associated visualfield loss but normal or low IOP. The nerve damage involves loss ofretinal ganglion cells in a characteristic pattern. There are manydifferent subtypes of glaucoma, but they can all be considered to be atype of optic neuropathy. Raised intraocular pressure (e.g., above 21mmHg or 2.8 kPa) is the most important and only modifiable risk factorfor glaucoma. However, some may have high eye pressure for years andnever develop damage, while others can develop nerve damage at arelatively low pressure. Untreated glaucoma can lead to permanent damageof the optic nerve and resultant visual field loss, which over time canprogress to blindness.

The term “uveitis” refers to an inflammatory disease of the uvea, thevascular layer of the eye sandwiched between the retina and the white ofthe eye (sclera). The uvea extends toward the front of the eye andconsists of the iris, choroid layer and ciliary body. Uveitis includesanterior uveitis, intermediate uveitis, and posterior uveitis. A mostcommon type of uveitis is an inflammation of the iris called iritis(anterior uveitis). Uveitis may also occur at the posterior segment ofthe eye (e.g., at the choroid). Inflammation of the uvea can berecurring and can cause serious problems such as blindness if leftuntreated (accounts for 10% of blindness globally). Early diagnosis andtreatment are important to prevent the complications of uveitis.

The term “dry eye” or “dry eyes” includes an ocular disease in whichthere is insufficient tears to lubricate and nourish the eye. Tears arenecessary for maintaining the health of the front surface of the eye andfor providing clear vision. Patients with dry eyes either do not produceenough tears or have a poor quality of tears. Dry eye is a common andoften chronic problem, particularly in older adults. With each blink ofthe eyelids, tears are spread across the front surface of the eye, knownas the cornea. Tears provide lubrication, reduce the risk of eyeinfection, wash away foreign matter in the eye, and keep the surface ofthe eyes smooth and clear. Excess tears in the eyes flow into smalldrainage ducts, in the inner corners of the eyelids, which drain in theback of the nose. Tears are produced by several glands (e.g., lacrimalgland) in and around the eyelids. Tear production tends to diminish withage, with various medical conditions, or as a side effect of certainmedicines. Environmental conditions such as wind and dry climates canalso affect tear volume by increasing tear evaporation. When the normalamount of tear production decreases or tears evaporate too quickly fromthe eyes, symptoms of dry eye can develop. The most common form of dryeyes is due to an inadequate amount of the water layer of tears. Thiscondition, called keratoconjunctivitis sicca (KCS), is also referred toas “dry eye syndrome.”

The term “diabetic retinopathy” includes retinopathy (i.e., a disease ofthe retina) caused by complications of diabetes, which can eventuallylead to blindness. Diabetic retinopathy may cause no symptoms, mildvision problems, or even blindness. Diabetic retinopathy is the resultof microvascular retinal changes. Hyperglycemia-induced intramuralpericyte death and thickening of the basement membrane lead toincompetence of the vascular walls. These damages change the formationof the blood-retinal barrier and also make the retinal blood vesselsbecome more permeable. The pericyte death is caused when hyperglycemiapersistently activates protein kinase C-δ (PKC-δ, encoded by Prkcd) andp38 mitogen-activated protein kinase (MAPK) to increase the expressionof a previously unknown target of PKC-δ signaling, Src homology-2domain—containing phosphatase-1 (SHP-1), a protein tyrosine phosphatase.This signaling cascade leads to PDGF receptor-dephosphorylation and areduction in downstream signaling from this receptor, resulting inpericyte apoptosis. Small blood vessels, such as those in the eye, areespecially vulnerable to poor control over blood sugar. Anoveraccumulation of glucose and/or fructose damages the tiny bloodvessels in the retina. During the initial stage, called“nonproliferative diabetic retinopathy” (NPDR), most patients do notnotice any change in their vision. Early changes that are reversible anddo not threaten central vision are sometimes termed simplex retinopathyor background retinopathy. As the disease progresses, severenonproliferative diabetic retinopathy enters an advanced, “proliferativediabetic retinopathy” (PDR) stage when blood vessels proliferate. Thelack of oxygen in the retina causes fragile, new, blood vessels to growalong the retina and in the clear, gel-like vitreous humor that fillsthe inside of the eye, which may result in bleeding, cloudy vision,retina damage, or tractional retinal detachment.

Treatment or prevention of a disease may be accomplished byadministering Compound 1 to a mammal, such as a human being, in needthereof.

Pharmaceutical Compositions

A crystalline form of Compound 1, such as a crystalline form E or acrystalline form F, hereafter referred to as a “crystalline form,” maybe present in a pharmaceutical composition that may further comprise apharmaceutically acceptable excipient.

A crystalline form of Compound 1 may be intended for delivery in asubject.

Excipients

A pharmaceutically acceptable excipient or pharmaceutically acceptablecarrier may include a non-toxic, inert solid, semi-solid or liquidfiller, diluent, encapsulating material or formulation auxiliary of anysuitable type. Any pharmaceutically acceptable excipient may be usedherein. Some examples of materials which can serve as pharmaceuticallyacceptable carriers are sugars such as lactose, glucose, and sucrose;starches such as corn starch and potato starch; cellulose and itsderivatives such as sodium carboxymethyl cellulose, ethyl cellulose, andcellulose acetate; powdered tragacanth; malt; gelatin; talc; excipientssuch as cocoa butter and suppository waxes; oils such as peanut oil,cottonseed oil; safflower oil; sesame oil; olive oil; corn oil andsoybean oil; glycols such as propylene glycol; esters such as ethyloleate and ethyl laurate; agar; detergents such as TWEEN 80; bufferingagents such as magnesium hydroxide and aluminum hydroxide; alginic acid;pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol;and phosphate buffer solutions, as well as other non-toxic compatiblelubricants such as sodium lauryl sulfate and magnesium stearate, as wellas coloring agents, releasing agents, coating agents, sweetening,flavoring and perfuming agents, preservatives and antioxidants can alsobe present in the composition, according to the judgment of theformulator. As would be appreciated by one of skill in this art, theexcipients may be chosen based on the route of administration asdescribed below, the pharmaceutical agent being delivered, time courseof delivery of the agent, etc.

Non-limiting examples of polymers which can be included in the disclosedpharmaceutical compositions include polyamines, polyethers, polyamides,polyesters, polycarbamates, polyureas, polycarbonates, polystyrenes,polyimides, polysulfones, polyurethanes, polyacetylenes, polyethylenes,polyethyeneimines, polyisocyanates, polyacrylates, polymethacrylates,polyacrylonitriles, and polyarylates. Non-limiting examples of specificpolymers include poly(caprolactone) (PCL), ethylene vinyl acetatepolymer (EVA), poly(lactic acid) (PLA), poly(L-lactic acid) (PLLA),poly(glycolic acid) (PGA), poly(lactic acid-co-glycolic acid) (PLGA),poly(L-lactic acid-co-glycolic acid) (PLLGA), poly(D,L-lactide) (PDLA),poly(L-lactide) (PLLA), poly(D,L-lactide-co-caprolactone),poly(D,L-lactide-co-caprolactone-co-glycolide), poly(D,L-lactide-co-PEO-co-D, L-lactide), poly(D, L-lactide-co-PPO-co-D,L-lactide), polyalkyl cyanoacrylate, polyurethane, poly-L-lysine (PLL),hydroxypropyl methacrylate (HPMA), poly(ethylene glycol),poly-L-glutamic acid, poly(hydroxy acids), polyanhydrides,polyorthoesters, poly(ester amides), polyamides, poly(ester ethers),polycarbonates, polyalkylenes such as polyethylene and polypropylene,polyalkylene glycols such as poly(ethylene glycol) (PEG), polyalkyleneterephthalates such as poly(ethylene terephthalate), polyvinyl alcohols(PVA), polyvinyl ethers, polyvinyl esters such as poly(vinyl acetate),polyvinyl halides such as poly(vinyl chloride) (PVC),polyvinylpyrrolidone, polysiloxanes, polystyrene (PS), polyurethanes,derivatized celluloses such as alkyl celluloses, hydroxyalkylcelluloses, cellulose ethers, cellulose esters, nitro celluloses,hydroxypropylcellulose, carboxymethylcellulose, polymers of acrylicacids, such as poly(methyl(meth)acrylate) (PMMA),poly(ethyl(meth)acrylate), poly(butyl(meth)acrylate),poly(isobutyl(meth)acrylate), poly(hexyl(meth)acrylate),poly(isodecyl(meth)acrylate), poly(lauryl(meth)acrylate),poly(phenyl(meth)acrylate), poly(methyl acrylate), poly(isopropylacrylate), poly(isobutyl acrylate), poly(octadecyl acrylate) (jointlyreferred to herein as “polyacrylic acids”), and copolymers and mixturesthereof, polydioxanone and its copolymers, polyhydroxyalkanoates,polypropylene fumarate), polyoxymethylene, poloxamers,poly(ortho)esters, poly(butyric acid), poly(valeric acid),poly(lactide-co-caprolactone), and trimethylene carbonate.

Route of Administration

Pharmaceutical compositions containing the particles described hereinmay be administered to a subject via any route known in the art. Theseinclude, but are not limited to, oral, sublingual, nasal, injection(e.g., intravenous, intradermal, subcutaneous, intramuscular), rectal,vaginal, intraarterial, intracisternally, intraperitoneal, intravitreal,periocular, topical (e.g., ocular or dermal, such as by powders, creams,ointments, or drops), buccal, and inhalational administration. In someembodiments, compositions described herein may be administeredparenterally as injections (intravenous, intramuscular, orsubcutaneous), drop infusion preparations, or suppositories. The routeof administration and the effective dosage to achieve the desiredbiological effect may be determined by the agent being administered, thetarget organ, the preparation being administered, time course ofadministration, disease being treated, intended use, etc.

Moreover, the pharmaceutical compositions may be administeredparenterally as injections (intravenous, intramuscular, orsubcutaneous), drop infusion preparations, or suppositories. Forophthalmic applications, the pharmaceutical compositions may beadministered by injection (e.g., intraocular, conjunctival,subconjunctival, intrastromal, intravitreal, or intracameral), or by thelocal or ophthalmic mucous membrane route, the pharmaceuticalcompositions may be administered topically, such as solutions,suspensions (e.g., eye drops), gels, or ointments.

In some embodiments, an effective amount of Compound 1 in a crystallineform may vary from about 0.00001 mg/kg to about 100 mg/kg in one or moredose administrations for one or several days (depending on the mode ofadministration). In certain embodiments, the effective amount per dosevaries from about 0.00001 mg/kg to about 100 mg/kg, from about 0.0001mg/kg to about 20 mg/kg, from about 0.001 mg/kg to about 5 mg/kg, fromabout 0.01 mg/kg to about 0.5 mg/kg, and from about 0.1 mg/kg to about 2mg/kg, 0.5-5 mg/kg, 1-10 mg/kg, or any dose amount bounded by or betweenany of these values.

Examples Example 1. Synthesis of Compound 1

Compound 1 may be prepared by the method described in U.S. Pat. No.9,458,169, which is hereby incorporated by reference for its descriptionof the synthesis of Compound 1.

Example 2. Preparation of Amorphous Compound 1

The synthesized Compound 1 (6.25 g) was suspended in water (300 mL).Hydrochloric acid (1M, 20 mL) was added. All material dissolved to forma light yellow solution. The solution was filtered through a disk filter(1 μm) and placed under high vacuum with magnetic stirring for 10minutes. While stirring a saturated sodium bicarbonate (300 mL) wasadded over 10 minutes. The high vacuum was applied for additional 10minutes and the solution was stirred for 30 minutes under normalpressure. The solid precipitate was filtered off on a sintered glassfunnel (F porosity). The solid was washed with water (300 mL) and driedby passage of vacuum for 1 hour. The solid was further dried under highvacuum overnight. The material obtained was a white amorphous solid(5.45 g).

Example 3. Preparation of Crystalline Form E

Crystalline form E was prepared by stirring a slurry of amorphousCompound 1 in methanol.

In one instance, amorphous Compound 1 (30.8 mg) was added into a 4-mLscintillation vial with screw-top. To the vial was added a magnetic stirbar and methanol (500 μL). The vial was capped and the suspension in thevial was allowed to stir at ambient temperature (about 22° C.) with 300RPM speed for two hours.

The solid material was collected by centrifuge filtration and thefiltrate was discarded. The centrifuge filter tube was then lightlycovered and dried under high vacuum for approximately 17 hours. Therecovered dry solid (26.7 mg) was analyzed by X-ray powder diffraction(XRPD), which showed a unique powder pattern that was assigned ascrystalline form E. This experiment was successfully reproduced threetimes as indicated in Table 1.

TABLE 1 Summary of conversion to crystalline form E from amorphous.Amorphous Volume of Initial Crystalline initially methanol Concen- FormE Recovery Experi- used added tration Recovered Yield by ment (mg) (uL)(mg/mL) (mg) mass 1 30.8 0.5 61.6 26.7 92.2% 2 29.6 0.5 59.2 25.5 91.6%3 29.7 0.5 59.4 26.6 95.2%

Properties of Crystalline Form E

The XRPD pattern of crystalline form E is illustrated in FIG. 1. Thepeaks in degrees 2θ, the corresponding d-spacing values, and relativeintensity (%) from the XRPD pattern of crystalline form E are listed inTable 2.

TABLE 2 XRPD peak listing of crystalline form E. Relative Peak Position± d-spacing ± Intensity No. 0.2° [2 θ] 0.2 [Å] [%] 1 8.55 10.33 12.85 28.95 9.87 8.72 3 9.43 9.37 6.27 4 10.22 8.65 100.00 5 14.19 6.23 14.09 614.65 6.04 9.27 7 14.90 5.94 28.70 8 15.38 5.76 40.99 9 15.86 5.58 52.7910 16.50 5.37 32.26 11 17.07 5.19 3.65 12 17.69 5.01 16.53 13 18.90 4.6945.03 14 19.43 4.57 35.00 15 20.33 4.36 12.97 16 20.57 4.31 39.09 1721.36 4.16 5.34 18 22.36 3.97 35.40 19 23.24 3.82 11.49 20 23.46 3.7967.26 21 23.75 3.74 45.38 22 25.63 3.47 26.16 23 26.76 3.33 8.83 2428.52 3.13 22.86 25 28.93 3.08 10.61 26 29.56 3.02 5.47 27 34.19 2.623.74

Thermogravimetric Analysis (TGA) was found to exhibit a mass loss of8.88%. Since solution ¹H NMR confirmed the presence of methanol in neatcrystalline form E, the TGA data suggests that the mass loss is possiblydue to loss of methanol.

Example 4. Preparation of Crystalline Form F

Crystalline form F was prepared by crystallization of amorphous Compound1 from a mixture of water and isopropanol (H₂O:IPA=1:9).

In one instance, amorphous Compound 1 (15.0 mg) was added into a 4-mLscintillation vial with screw-top. To the vial was added a magnetic stirbar and a mixture of water and isopropanol (H₂O;IPA=1:9, 1 mL). The vialwas capped and placed onto a hot plate equipped with a magnetic stiringspeed control that was preheated to 50° C. The vial was allowed to stirat 50° C. with 400 RPM speed for one hour to fully dissolve Compound 1.Following dissolution, the heating and stirring were halted, and thestirring bar was removed from the hot solution using a magneticretriever. The vial was recapped then removed from the hot plate and wasstored under ambient conditions (approximately 22° C.) for 17 hours.During this period, the solution spontaneously cooled to temperature(approximately 22° C.).

After incubation at room temperature, crystals were visibly present inthe vial. The crystalline material was collected by centrifugefiltration and the filtrate was discarded. The centrifuge filter tubewas lightly covered then dried under high vacuum for approximately 17hours. The recovered dry solid (8.4 mg) was analyzed by XRPD, whichshowed a unique powder pattern that was assigned as crystalline form F.This experiment was successfully reproduced three times as indicated inTable 3.

TABLE 3 Summary of crystallization to produce crystalline form F fromamorphous Compound 1. Volume Amorphous of 1:9 Initial Crystallineinitially water:IPA Concen- form F Recovery Experi- used added trationRecovered Yield by ment (mg) (mL) (mg/mL) (mg) mass 1 15.0 1 15.0 8.459.98% 2 15.8 1 15.8 8.4 56.94% 3 15.1 1 15.1 9.5 67.38%

Crystalline form F was also prepared by crystallization of crystallineform E of Compound 1 from of water and isopropanol (H₂O:IPA=1:9).

In one instance, crystalline form E of Compound 1 (15.5 mg) was addedinto a 4-mL scintillation vial with screw-top. To the vial was added amagnetic stirring bar and a mixture of water and isopropanol(H₂O:IPA=1:9, 1 mL). The vial was capped and placed onto a hot plateequipped with a magnetic stirring speed control that was preheated to50° C. The vial was allowed to stir at 50° C. with 400 RPM speed for onehour to fully dissolve Compound 1. Following dissolution, the heatingand stirring were halted, and the stir bar was removed from the hotsolution using a magnetic retriever. The vial was recapped then removedfrom the hot plate and was stored under ambient conditions for 17 hours.During this period, the solution spontaneously cooled to temperature(approximately 22° C.).

After cooled down to room temperature, crystals were visibly present inthe vial. The mother liquor was decanted. The vial containing solid waslightly covered, then dried under high vacuum overnight. The recovereddry solid (12.3 mg) was analyzed by XRPD on the next day, which showed aunique powder pattern that was assigned as crystalline form F. Thisexperiment was successfully reproduced three times as indicated in

Table 4.

TABLE 4 Summary of crystallization to produce crystalline form F fromcrystalline form E. Volume Crystalline of 1:9 of Crystalline form Ewater:IPA Initial form F Recovery initially used added ConcentrationRecovered Yield by Experiment (mg) (mL) (mg/mL) (mg) mass 1 15.3 1 15.3Not Not determined determined 2 15.5 1 15.5 12.3 78.1% 3 15.1 1 15.113.4 88.1%

Properties of Crystalline Form F

The XRPD pattern of crystalline form F is illustrated in FIG. 2. Thepeaks in degrees 2θ, the corresponding d-spacing values, and relativeintensity (%) in the XRPD pattern of crystalline form F are listed inTable 5.

TABLE 5 XRPD peak listing of neat crystalline form F. Relative PeakPosition ± d-spacing ± Intensity No. 0.2° [2 θ] 0.2 [Å] [%] 1 7.74 11.4239.6 2 9.60 9.21 13.1 3 12.94 6.84 79.1 4 13.69 6.47 17.3 5 14.63 6.0519.6 6 14.87 5.95 45.9 7 15.55 5.69 11.7 8 17.65 5.02 100.0 9 18.27 4.853.5 10 19.85 4.47 5.5 11 20.46 4.34 5.6 12 20.66 4.30 7.2 13 22.20 4.0020.1 14 22.68 3.92 15.6 15 22.97 3.87 29.8 16 23.18 3.83 38.9 17 25.163.54 3.8 18 26.62 3.35 88.1 19 27.41 3.25 12.1

TGA was found to exhibit a total mass loss of 5.2%. Since solution ¹HNMR showed insignificant amounts of isopropanol in neat crystal form F,the mass loss in TGA indicates that crystalline form F is possibly ahydrate.

The foregoing description details specific methods and compositions thatcan be employed to make and use the compounds described herein, andrepresents the best mode contemplated. However, it is apparent for oneof ordinary skill in the art that other compounds with the desiredpharmacological properties can be prepared in an analogous manner, andthat the disclosed compounds can also be obtained from differentstarting compounds via different chemical reactions. Similarly,different pharmaceutical compositions may be prepared and used withsubstantially the same result. Thus, the foregoing description shouldnot be construed as limiting the scope of the claims.

Unless otherwise indicated, all numbers expressing quantities ofingredients, properties such as molecular weight, reaction conditions,and so forth used in the specification and claims are to be understoodas being modified in all instances by the term “about.” Accordingly,unless indicated to the contrary, the numerical parameters set forth inthe specification and attached claims are approximations that may varydepending upon the desired properties sought to be obtained by thepresent invention. At the very least, and not as an attempt to limit theapplication of the doctrine of equivalents to the scope of the claims,each numerical parameter should at least be construed in light of thenumber of reported significant digits and by applying ordinary roundingtechniques. Notwithstanding that the numerical ranges and parameterssetting forth the broad scope of the invention are approximations, thenumerical values set forth in the specific examples are reported asprecisely as possible. Any numerical value, however, inherently containscertain errors necessarily resulting from the standard deviation foundin their respective testing measurements.

The terms “a,” “an,” “the” and similar referents used in the context ofdescribing the invention (especially in the context of the followingclaims) are to be construed to cover both the singular and the plural,unless otherwise indicated herein or clearly contradicted by context.Recitation of ranges of values herein is merely intended to serve as ashorthand method of referring individually to each separate valuefalling within the range. Unless otherwise indicated herein, eachindividual value is incorporated into the specification as if it wereindividually recited herein. All methods described herein can beperformed in any suitable order unless otherwise indicated herein orotherwise clearly contradicted by context. The use of any and allexamples, or exemplary language (e.g., “such as”) provided herein isintended merely to better illuminate the invention and does not pose alimitation on the scope of the invention otherwise claimed. No languagein the specification should be construed as indicating any non-claimedelement essential to the practice of the invention.

Groupings of alternative elements or embodiments of the inventiondisclosed herein are not to be construed as limitations. Each groupmember may be referred to and claimed individually or in any combinationwith other members of the group or other elements found herein. It isanticipated that one or more members of a group may be included in, ordeleted from, a group for reasons of convenience and/or patentability.When any such inclusion or deletion occurs, the specification is deemedto contain the group as modified thus fulfilling the written descriptionof all Markush groups used in the appended claims.

Certain embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention. Ofcourse, variations on these described embodiments will become apparentto those of ordinary skill in the art upon reading the foregoingdescription. The inventor expects skilled artisans to employ suchvariations as appropriate, and the inventors intend for the invention tobe practiced otherwise than specifically described herein. Accordingly,this invention includes all modifications and equivalents of the subjectmatter recited in the claims appended hereto as permitted by applicablelaw. Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the invention unlessotherwise indicated herein or otherwise clearly contradicted by context.

Specific embodiments disclosed herein may be further limited in theclaims using consisting of or consisting essentially of language. Whenused in the claims, whether as filed or added per amendment, thetransition term “consisting of” excludes any element, step, oringredient not specified in the claims. The transition term “consistingessentially of” limits the scope of a claim to the specified materialsor steps and those that do not materially affect the basic and novelcharacteristic(s). Embodiments of the invention so claimed areinherently or expressly described and enabled herein.

Furthermore, numerous references have been made to patents and printedpublications throughout this specification. Each of the above-citedreferences and printed publications are individually incorporated hereinby reference in their entirety.

In closing, it is to be understood that the embodiments disclosed hereinare illustrative of the principles of the present invention. Othermodifications that may be employed are within the scope of theinvention. Thus, by way of example, but not of limitation, alternativeconfigurations of the present invention may be utilized in accordancewith the teachings herein. Accordingly, the present invention is notlimited to that precisely as shown and described.

What is claimed is:
 1. A method of inhibiting aberrant signaling of agrowth factor comprising administering to a subject a therapeuticallyeffective amount of a crystalline form of Compound 1:

wherein the crystalline form has an XRPD pattern comprising peaks at 7-8degrees 2θ, 12.5-13.5 degrees 2θ, 14.5-15.5 degrees 2θ, 17-18 degrees2θ, and 26-27 degrees 2θ, or a pharmaceutical composition comprising thecrystalline form.
 2. The method of claim 1, wherein the growth factor isvascular endothelial growth factor.
 3. The method of claim 2, whereinaberrant signaling of a growth factor is associated with a diseaseselected from cancer, a benign neoplasm, atherosclerosis, hypertension,an inflammatory disease, rheumatoid arthritis, and an ocular disease. 4.The method of claim 1, wherein the crystalline form is a hydrate.
 5. Themethod of claim 1, wherein the pharmaceutical composition is suitablefor topical administration.
 6. The method of claim 3, wherein the oculardisease is macular edema.
 7. The method of claim 6, wherein the oculardisease is a diabetic macular edema or cystoid macular edema.
 8. Themethod of claim 7, wherein the ocular disease is a macular degeneration.9. The method of claim 3, wherein the ocular disease is a retinal veinocclusion.
 10. The method of claim 3, wherein the ocular disease isretinopathy.
 11. The method of claim 8, wherein the ocular disease isage-related macular degeneration (AMD).
 12. The method of claim 3,wherein the ocular disease is corneal neovascularization.
 13. The methodof claim 3, wherein the ocular disease is glaucoma.
 14. The method ofclaim 3, wherein the ocular disease is inflammatory meibomian glanddisease.
 15. The method of claim 3, wherein the ocular disease is dryeye syndrome.
 16. The method of claim 3, wherein the ocular disease isblepharitis.
 17. The method of claim 3, wherein the ocular disease isuveitis.
 18. The method of claim 3, wherein the ocular disease isallergic conjunctivitis.
 19. The method of claim 3, wherein the oculardisease is rosacea of the eye.
 20. A kit comprising a crystalline formof Compound 1:

wherein the crystalline form has an X-ray powder diffraction (XRPD)pattern comprising peaks at 7-8 degrees 2θ, 12.5-13.5 degrees 2θ,14.5-15.5 degrees 2θ, 17-18 degrees 2θ, and 26-27 degrees 2θ, or apharmaceutical composition comprising the crystalline form.